Evaluation of sonic hedgehog, sex-determining region Y-box transcription factor 3 and wingless-type family member 3A and 9B in different cleft tissue

Cleft lip and palate pathogenesis has been characterized by the disrupted function of multiple regulatory genes and their coded proteins during the formation and fusion of facial folds. Multiple signaling pathways regulate facial region development and involve multiple regulatory proteins. The role of some of these proteins during cleft formation process is still uncertain in human tissue.
Our research analyses regulatory proteins Sonic Hedgehog, Sex-determining Region Y-Box Transcription Factor 3, and Wingless-type Family Member 3A and 9B proteins in the epithelium and connective tissue of different non-syndromic clefts by applying immunohistochemistry. Cleft tissue obtained during cleft-correcting surgery was divided into 3 patient groups based on cleft diagnosis – unilateral cleft lip patients with 36 individuals, bilateral cleft lip patients with 13 individuals and cleft palate patients with 26 individuals. The control group was made from lip tissue taken from 5 individuals not affected by clefts. The semiquantitative counting method was used for slide evaluation. Non-parametric statistics (Kruskal–Wallis H test, Mann-Whitney U test, Spearman’s correlation coefficient calculation) were used for result evaluation.
The results of our research showed that Sonic Hedgehog protein immunoreactive cells were significantly reduced in bilateral cleft lip and cleft palate tissue in comparison to controls. Sex-determining Region Y-Box Transcription Factor 3, Wingless-type Family Member 3A and Wingless-type Family Member 9B immunoreactive cells were notably decreased in number in all three cleft tissue types in comparison to controls. Multiple similar statistically notable correlations between the number of protein-containing cells were detected.
It was concluded that the notable decrease of Sonic Hedgehog immunoreactive cells in bilateral cleft lip and cleft palate tissue could associate this protein more strongly with the morphopathogenesis of these non-syndromic orofacial cleft types. The significant decrease of immunoreactive cells containing Sex-determining Region Y-Box Transcription Factor 3, Wingless-type Family Member 3A and Wingless-type Family Member 9B, might be involved with the pathogenetic development of all three evaluated cleft tissue types. Similar correlations between protein-containing cells in all three cleft tissue types seem to indicate similarities in pathogenetic mechanisms while notable differences in correlations are also present.